Stationary phase cavity

The master retention equation of the solvation parameter model relating the above processes to experimentally quantifiable contributions from all possible intermolecular interactions was presented in section 1.4.3. The system constants in the model (see Eq. 1.7 or 1.7a) convey all information of the ability of the stationary phase to participate in solute-solvent intermolecular interactions. The r constant refers to the ability of the stationary phase to interact with solute n- or jr-electron pairs. The s constant establishes the ability of the stationary phase to take part in dipole-type interactions. The a constant is a measure of stationary phase hydrogen-bond basicity and the b constant stationary phase hydrogen-bond acidity. The / constant incorporates contributions from stationary phase cavity formation and solute-solvent dispersion interactions. The system constants for some common packed column stationary phases are summarized in Table 2.6 [68,81,103,104,113]. Further values for non-ionic stationary phases [114,115], liquid organic salts [68,116], cyclodextrins [117], and lanthanide chelates dissolved in a poly(dimethylsiloxane) [118] are summarized elsewhere. 

However, in LC solutes are partitioned according to a more complicated balance among various attractive forces solutes interact with both mobile-phase molecules and stationary-phase molecules (or stationary-phase pendant groups), the stationary-phase interacts with mobile-phase molecules, parts of the stationary phase may interact with each other, and mobile-phase molecules interact with each other. Cavity formation in the mobile phase, overcoming the attractive forces of the mobile-phase molecules for each other, is an important consideration in LC but not in GC. Therefore, even though LC and GC share a considerable amount of basic theory, the mechanisms are very different on a molecular level. This translates into conditions that are very different on a practical level so different, in fact, that separate instruments are required in modern practice. 

In relation to separation of nucleotides, Hoffman61 found that adenine nucleotides interacted most strongly with cycloheptaamylose, presumably by inclusion of the base within the cavity of cyclodextrin. When epichlorohydrin-cross-linked cycloheptaamylose gel was used as a stationary phase for nucleic acid chromatography, adenine-containing compounds were retarded most strongly. 

The stationary phase in gel permeation (also called size exclusion) chromatography contains cavities of a defined size distribution, called pores. Analytes larger than the pores are excluded from the pores and pass through the column more rapidly than smaller analytes. There may be secondary effects due to hydrophobic adsorption, ionic interaction, or other interactions between the stationary phase and analyte. Gel permeation and non-ideal interactions in gel permeation are described more fully in Chapter 6. 

The elution of [60]- and [70]fullerenes was measured in water-methanol as a function of temperature on a poly(octadecylsiloxane) phase.67 The retention was shown to be dependent on the surface tension of the stationary phase through a simple geometrical model in which the solute formed a cavity in the stationary phase. In affinity chromatography, it was demonstrated that low ligand density may be a requirement for specificity of binding.68... 

Analytical shape computation techniques were applied for the detection of cavities and the calculation of molecular surface properties of isolated cavity features and other ordered formations within these resultant alkyl stationary-phase simulation models [227]. Deep cavities (8-10 A wide) within the alkyl chains were identified for Cig polymeric models representing shape selective stationary phases (Figure 5.23). Similar-structure cavities with significant alkyl-chain ordered regions (>11 A) were isolated from two independent Cig models (differing in temperature,... 

FIGURE 5.26 (See color insert following page 280.) A representation of the slot model illustrating potential constrained-shape solute (BaP) interactions with the conformational ordered cavities of a polymeric Cjg stationary-phase simulation model. Also included on the chromatographic surface is an identical-scale molecular structure of 1,2 3,4 5,6 7,8-tetrabenzonaphthalene (TBN). 

The approach to calculate the van der Waals and cavity terms from the molecular surface areas can be used for the calculation of partition coefficients. The results show that for the distribution of hydrocarbons between water and n-octanol the calculated partition coefficient is linear in carbon number. Qualitatively similar data are obtained for the distribution between other solvents and water and the results can be used to predict the retention in liquid>liquid chromatography. On the other hand, if retention in RPC occurs due to reversible binding at the surface of the stationary phase, the significant parameter is not the total surface area of the eiuite but rather the net decrease in the molecular surface area of the stationary phase ligates and that of the eiuite upon binding, i.e., the contact area in the complex. 

Porosity is one of the most important properties of a stationary phase, since it severely inflnences the chromatographic colnmn performance, the speed of separation, as well as the specific surface area and consequently loading capacity. Porosity refers to the degree and distribution of the pore space present in a material [114], Open pores indicate cavities or channels, located on the surface of a particle, whereas closed pores are situated inside the material. The sum of those pores is defined as intraparticular porosity. Interparticular porosity, in contrast, is the sum of all void volume between the particles. According to their diameter, pores have been internationally (lUPAC) classified as follows [114] ... 

For example, cyclodextrins form chiral cavities which adsorb the corresponding enantiomers with different affinity while cellulose triacetate crystallizes in the form of helical substructures in which the enantiomers may be incorporated with different rates. For amino acid derived stationary phases there are two types of enantiomer differentiating interactions a brush-like hydrogen bond and dipole interaction plus a /[-complex donor or acceptor interaction with the aromatic residues in the amino acid. 

The term Molecular Clip has been coined for molecules of type 2. That these molecules do indeed possess the geometric features of a clip is apparent from the X-ray structure of the tetramethoxy derivative 3a (Fig. 2) [lla,b]. The o-xylylene moieties of this molecule define a tapering cavity, the walls of which are at an angle of 39.5 with the centers of the benzene rings 6.67 A apart. The carbonyl groups of the glycoluril moiety, which are hydrogen-bond acceptor sites, are separated by 5.52 A. It was also possible to obtain a crystal structure of the chiral dibromo-derivative 4 of clip 3 (Fig. 3). This compound was separated into its enantiomers by HPLC on a chiral stationary phase [12]. 

Many types of chiral stationary phase are available. Pirkle columns contain a silica support with bonded aminopropyl groups used to bind a derivative of D-phenyl-glycine. These phases are relatively unstable and the selectivity coefficient is close to one. More recently, chiral separations have been performed on optically active resins or cyclodextrins (oligosaccharides) bonded to silica gel through a small hydrocarbon chain linker (Fig. 3.11). These cyclodextrins possess an internal cavity that is hydro-phobic while the external part is hydrophilic. These molecules allow the selective inclusion of a great variety of compounds that can form diastereoisomers at the surface of the chiral phase leading to reversible complexes. 

Figure 3.11—Separation on a cyclodextrin-boimd stationary phase. Chromatogram of a racemic mixture chemical formula of /f-cyclodextrin (diameter, 1.5 nm cavity, 0.8 nm height, 0.8 nm) partial representation of cyclodextrin bonded to a silica gel bead through an alkyl chain linker arm side view of a cyclodextrin molecule with a hydrophobic cavity.

Radius of holiow cavity r, = 124 pm Radius to middle of stationary phase r2 = 124.5 pm... 

Common chiral stationary phases for gas chromatography have cyclodextrins bonded to a conventional polysiloxane stationary phase.7-8 Cyclodextrins are naturally occurring cyclic sugars. P-Cyclodextrin has a 0.78-nm-diameter opening into a chiral, hydrophobic cavity. The hydroxyls are capped with alkyl groups to decrease the polarity of the faces.9... 

Generally, CD-based chiral stationary phases have been used in the reversed-phase mode. Earlier, it was assumed that in the normal phase mode, the more nonpolar component of the mobile phase would occupy the CD cavity, thereby blocking inclusion complexation between the chiral analyte and CD [4,11], But with the development of CD derivatives, it has become possible to use the normal phase mode too [45,74], Among the various CSPs based on CD derivatives, one based on a naphthylethyl carbamoylated derivative has shown excellent enantioselectivity in the normal phase mode [46,59]. Armstrong et al. [45] synthesized several /CCD derivatives and had them tested in the normal phase mode to resolve the enantiomers of a variety of drugs hexane-2-propanol (90 10, v/v) served as the mobile phase. The authors discussed the similarities and differences of the enantioselectivities on the native and derivatized CD phases. 

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